Industrial fermentation

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Darrell Cook
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Industrial fermentation Production with microbes Industrial microbiology The use of microbes

beverage industry, vinegar, baking yeast, citric and lactic acid Late 20 th century

Fermentation industry Ability of microbes to produce useful product Usually a naturally

Metabolites Primary metabolite produce during growth Secondary metabolite produce after

Product Substrate Trophophase Idiophase Time Production with microbes Interdisciplinary area

.of course Molecular biologists Chemists/biochemists Engineers hardware, software Lawyers

1 Industrial fermentation Production with microbes Industrial microbiology The use of microbes in the production Start from an ancient time Large scale production Early 20 th century beverage industry, vinegar, baking yeast, citric and lactic acid Late 20 th century development of biotechnology from the antibiotic industrial Production with microbes Fermentation industry Ability of microbes to produce useful product Usually a naturally produce substances for growth, and maintenance. Metabolites Primary metabolite produce during growth Secondary metabolite produce after growth stage Cell Product Substrate Time Primary metabolite Secondary metabolite Cell Product Substrate Trophophase Idiophase Time Production with microbes Interdisciplinary area Microbiologists..of course Molecular biologists Chemists/biochemists Engineers hardware, software Lawyers patent and permission Investors Management team! Products I. Foods and foods related - Fermented meat, Cheeses and milk products, mushrooms, baker s s yeast, coffee, pickles, single cell-proteins, vinegar, amino acids, vitamins, alcoholic beverages 1

Enzymes and microbial transformation - Commercial

Engineered microbes - Insulin, human growth factor

fermentation Isolation of microbes that produce your

lab Scale up from lab scale (up to 10 L) to

) Whole cells products Including mushrooms and

mold! Highly consumed every year high nutrition.

Stalk and cap = compact mycelium Gills = site of

Agaricus bisporus (botton mushroom), Lentinus edulus

protein yeast, and cyanobacteria Alternative food

2 Products II. Organic acids - Citric acids, Itaconic acid III. Enzymes and microbial transformation - Commercial enzyme, sterol conversion IV. Engineered microbes - Insulin, human growth factor Production with microbes Steps in Industrial fermentation Isolation of microbes that produce your interested product Screen for the best producing strain: naturally or mutation. Optimize production condition (growth basically) in lab Scale up from lab scale (up to 10 L) to industrail scale (>10,000 L) (raw material?) Whole cells products Including mushrooms and Single-cell protein (SCP) Mushrooms fruiting body of mold! Highly consumed every year high nutrition. Grow on decaying organic matter in soil or wood. Stalk and cap = compact mycelium Gills = site of reproductive spores E.g. Agaricus bisporus (botton mushroom), Lentinus edulus (Shitake mushroom) Agaricus bisporus Single-cell protein yeast, and cyanobacteria Alternative food sources mostly used now as supplementary diet e.g. yeast is a source for B Vitamins. Organisms: yeast, Spirulina Growing biomass and freeze dry Raw material from other industrial, farming Shitake mushroom Oyster mushroom Straw mushroom 2

3 Baker s s yeast Same process as in single cell protein Cheese production Can produce from any milk. 4 phases: coagulation, separation, shaping and ripening Coagulation: rennin enzyme or acids precipitate casein From calves s s stomachs, engineered bacteria Acid production by lactic acid bacteria Curd entrap bacteria, fat globules and other material. Cheese production Additives (salt and herbs) may be added after seperation or shaping. Additives and shape effect property of cheese product Ripening add flavor and look Natural or inoculation of microbes e.g. Penicillium roquefortii (spores( are usually added into blue cheese), and Penicillium camemberti is added to the surface of a cheese known as Camenbert Blue cheese Brie Gorgonzola Swiss 3

Yogurt and other fermented milk products Yogurt,

acid production Lactobacillus bulgaricus 0.5-1.

acetaldehyde, diacetyl, and acetoin Product need

cream Cultured buttermilk Acidophilus milk Kefir

Microorganisms Streptococcus sp. Leuconostoc sp.

Lactobacillus acidophilus Streptococcus lactis,

Cream is inoculated and leave to develop acidity

milk. This milk product is used for its medicinal

A mixed lactic acid and alcoholic fermentation.

Alcoholic beverages Accidental discovered from

Fermentation of sugar in fruit, grains and other

polysaccharide to fermentable sugar Raw materials

4 Yogurt and other fermented milk products Yogurt, kefir, buttermilk, sourcream Yogurt usually use mixture of 2 cultures: Streptococcus thermophilus: : ini. acid production Lactobacillus bulgaricus % lactic acid Fermented at C Taste due to acetaldehyde, diacetyl, and acetoin Product need to contains live cultures Fermented Product Sour cream Cultured buttermilk Acidophilus milk Kefir Some Foods Produced from Fermented Milk. Microorganisms Streptococcus sp. Leuconostoc sp. Streptococcus sp. Leuconostoc sp. Lactobacillus acidophilus Streptococcus lactis, Lactobacillus bulgaricus, yeasts Description Cream is inoculated and leave to develop acidity Made with skimmed or partly skimmed pasteurized milk. This milk product is used for its medicinal therapeutic value. A mixed lactic acid and alcoholic fermentation. Industrial production of yogurt Kefir Buttermilk Alcoholic beverages Accidental discovered from ancient human Saccharomyces cerevisiae Fermentation of sugar in fruit, grains and other In case of other materials need to convert polysaccharide to fermentable sugar Raw materials are regional.variety of products Wine grapes, fruits Beer cereal grain barley, sorgum, wheat Other rice wine (rice), teguila (agave cactus), vodka (corn), mead (honey). Tequila 4

5 Fermented meat Use to preserve food E.g. salami, summer sausage, cured ham Thailand naam (fermented pork), fermented sausage, fermented fish, fish sauce etc etc Main organism Pediococcus serevisiae, Lactobacillus plantarum, other lactic acid bacteria Production of lactic acid to change ph of food Production of probiotics in some strains Fermented fruits and vegetables Found around the world Hundreds if not thousand type of products With bacteria, yeast and mold Depends mainly on native microbes associate with the plants. Most included mixed culture fermentation Conditioning by human to favor growth of beneficial microbes - salting Products Sauerkraut Pickles Soy sauce Coffee Cocoa products Organisms Lactobacillus mesenteroids, Lactobacillus plantarum Pediococcus cerevisiae, Lactobacillus plantarum Rhizopus oligosporus, Rhizopus oryzae, Lactobacillus delbrueckii, Saccharomyces rouxii Leuconostoc mesenteroides, Saccharomyces marscianus, Flavobacterium spp., Fusarium spp. Leuconostoc mesenteroides, Candida sp. Flavoring agents: amino acids and nucleotides Monosodium glutamate, inosinic acid flavor enhancers Originally use seaweed, fish Produce by enzymatic hydrolysis of yeast RNA (Candida utilis) Now direct fermentation = monosodium glutamate and 5 -IMP 5 (inosine monophosphate) Brevibacterium ammoniagenes produce inosine and then chemically phosphorylated Amino acids Sodium glutamate L-lysine L-methionine L-threonine L-tryptophane L-tryptophane L-histidine Aspartate Alanine L-aspartyl-l-phenylalanine Methyl ester (aspartame) Mixed amino acid Utilization Flavor enhancer Added to plant derived food to inprove nutritional value Antioxidant in powdered milk Improve flavor of fruit juices Low-caloric sweetener Infusion solutions for surgery patients 5

Amino acid L-glutamic L-lysine Organism Corynebacterium glutamicum Corynebacterium Approx.

Enzyme Bacterial proteases Asparaginase Glucoamylase Bacterial amylases Glucose isomerase Escherichia, Serratia Aspergillus Bacillus

baking, feed, detergents Sweeteners Use Rennin Pectinase Alcaligenes, Aspergillus, Candida Aspergillus Cheese manufacture Fruit juice

Antibiotics 3. Alkaloids 4. Steroids 5. Non-Steroid Hormones/cell regulators (cytokines): 1. Epidermal growth factor 2. Proinsulin 3.

6 Amino acid L-glutamic L-lysine Organism Corynebacterium glutamicum Corynebacterium Approx. yield (g/l) > C-source Glucose Glucose L-lysine L-threonine Brevibacterium flavum Escherichia coli K Acetate Sucrose Corynebacterium glutamicum E. coli Non-food products Enzyme production Produce in large scale around the world Major producer Novozyme (44%), Genencor Intl, USA (18%) Enzyme Bacterial proteases Asparaginase Glucoamylase Bacterial amylases Glucose isomerase Escherichia, Serratia Aspergillus Bacillus Organisms Bacillus, Streptomyces Bacillus, Streptomyces Detergents Antitumor agent Fructose syrup production Starch liquefaction, brewing, baking, feed, detergents Sweeteners Use Rennin Pectinase Alcaligenes, Aspergillus, Candida Aspergillus Cheese manufacture Fruit juice clarification Lipases Micrococcus Cheese production Penicillin acylase Escherichia Semisynthetic penicillins Non-food product 1. Vitamins 2. Antibiotics 3. Alkaloids 4. Steroids 5. Non-Steroid Hormones/cell regulators (cytokines): 1. Epidermal growth factor 2. Proinsulin 3. Insulin 4. Human growth hormone 5. Somatostatin 6. Interferons 7. Platelet-derived growth factor 8. Fibroblast growth factor 9. Tumor Necrosis Factor 10. Other cytokines are coming on line all the time Non-food product 6. Blood coagulating factor XIII 7. The restriction enzymes 8. Other enzymes (e.g. ligase, DNA polymerases etc.) 9. Solvents 6

How do the microbes produce these products?

that encode protein of interest into vectors and

Able to produce high amount of compounds than in

Microbes able to utilize waste from other process

Antibiotics Penicillin by Penicillium chrysogenum

soil microorganisms especially in Streptomyces spp.

7 How do the microbes produce these products? Genetically modified microorganisms Clone genes that encode protein of interest into vectors and express this genes in microbes used in production. Able to produce high amount of compounds than in original organisms with ease of manipulation. Microbes able to utilize waste from other process as a raw material for fermentation. Antibiotics Penicillin by Penicillium chrysogenum Now most of the antibiotics were discover from soil microorganisms especially in Streptomyces spp. Engineered genes into production strains. Modification of existing antibiotics. 7

Alkaloids Nitrogen containing organic compounds that derived from plants Have medicinal purposes:

local anesthetic Quinine, caffeine, nicotine, strychnine, serotonin, and LSD Streptomycin from

hormones Human proteins Biotech Drugs Mainly recombinants proteins and monoclonal antibodies.

8 Alkaloids Nitrogen containing organic compounds that derived from plants Have medicinal purposes: Atropine dilates pupils of the eyes (some muscle disease) Morphine and codeine relief of pain Cocaine local anesthetic Quinine, caffeine, nicotine, strychnine, serotonin, and LSD Streptomycin from Streptomyces griseus serotonin Atropine Steroids Types of products include cortisone and reproductive hormones Human proteins Biotech Drugs Mainly recombinants proteins and monoclonal antibodies. Highly active fields of biotech Lots of new products each year For list of product approve in US Timeline in drug development 8

Applied and Industrial Microbiology

11/28/2016 PowerPoint Lecture Presentations prepared by Bradley W. Christian, McLennan Community College CHAPTER 28 Applied and Industrial Microbiology Industrial Food Canning Mesophilic bacteria can spoil